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At least 127 records · Page 7

Degradation Reduces Microbial Richness and Alters Microbial Functions in an Australian Peatland

Abstract Peatland ecosystems cover only 3% of the world’s land area; however, they store one-third of the global soil carbon (C). Microbial communities are the main drivers of C decomposition in peatlands, yet we have limited knowledge of their structure and function. While the microbial communities in the Northern Hemisphere peatlands are well documented, we have limited understanding of microbial community composition and function in the Southern Hemisphere peatlands, especially in Australia. We investigated the vertical stratification of prokaryote and fungal communities from Wellington Plains peatland in the Australian Alps. Within the peatland complex, bog peat was sampled from the intact peatland and dried peat from the degraded peatland along a vertical soil depth gradient (i.e., acrotelm, mesotelm, and catotelm). We analyzed the prokaryote and fungal community structure, predicted functional profiles of prokaryotes using PICRUSt, and assigned soil fungal guilds using FUNGuild. We found that the structure and function of prokaryotes were vertically stratified in the intact bog. Soil carbon, manganese, nitrogen, lead, and sodium content best explained the prokaryote composition. Prokaryote richness was significantly higher in the intact bog acrotelm compared to degraded bog acrotelm. Fungal composition remained similar across the soil depth gradient; however, there was a considerable increase in saprotroph abundance and decrease in endophyte abundance along the vertical soil depth gradient. The abundance of saprotrophs and plant pathogens was two-fold higher in the degraded bog acrotelm. Soil manganese and nitrogen content, electrical conductivity, and water table level (cm) best explained the fungal composition. Our results demonstrate that both fungal and prokaryote communities are shaped by soil abiotic factors and that peatland degradation reduces microbial richness and alters microbial functions. Thus, current and future changes to the environmental conditions in these peatlands may lead to altered microbial community structures and associated functions which may have implications for broader ecosystem function changes in peatlands.

Environmental Sciences & Ecology↗

Auriga Streams – I: disrupting satellites surrounding Milky Way-mass haloes at multiple resolutions

In a hierarchically formed Universe, galaxies accrete smaller systems that tidally disrupt as they evolve in the host’s potential. We present a complete catalogue of disrupting galaxies accreted onto Milky Way-mass haloes from the Auriga suite of cosmological magnetohydrodynamic zoom-in simulations. We classify accretion events as intact satellites, stellar streams, or phase-mixed systems based on automated criteria calibrated to a visually classified sample, and match accretions to their counterparts in haloes re-simulated at higher resolution. Most satellites at the present day have lost substantial amounts of stellar mass – 67 per cent have $f_\text{bound} < 0.97$ (our threshold of lost stellar mass to no longer be considered intact), while 53 per cent satisfy a more stringent $f_\text{bound} < 0.8$. Streams typically outnumber intact systems, contribute a smaller fraction of overall accreted stars, and are substantial contributors at intermediate distances from the host centre ($\sim$0.1 to $\sim 0.7R_\text{200m}$, or $\sim$35 to $\sim$250 kpc for the Milky Way). We also identify accretion events that disrupt to form streams around massive intact satellites instead of the main host. Streams are more likely than intact or phase-mixed systems to have experienced pre-processing, suggesting this mechanism is important for setting disruption rates around Milky Way-mass haloes. All of these results are preserved across different simulation resolutions, though we do find some hints that satellites disrupt more readily at lower resolution. The Auriga haloes suggest that disrupting satellites surrounding Milky Way-mass galaxies are the norm and that a wealth of tidal features waits to be uncovered in upcoming surveys.

galaxies: haloes↗

Tree Mortality Decreases Water Availability and Ecosystem Resilience to Drought in Piñon-Juniper Woodlands in the Southwestern U.S.: Tree Mortality in Semiarid Biomes

Climate-driven tree mortality has increased globally in response to warmer temperature and more severe drought. To examine how tree mortality in semi-arid biomes impacts surface water balance, we experimentally manipulated a piñon-juniper (PJ) woodland by girdling all adult piñon trees in a 4 ha area, decreasing piñon basal area by ~65%. Over 3.5 years (2009-2013), we compared water flux measurements from this girdled site with those from a nearby intact PJ woodland. Before and after girdling, the ratio of evapotranspiration (ET) to incoming precipitation was similar between the two sites. Girdling altered the partitioning of ET such that the contribution of canopy transpiration to ET decreased 9-14% over the study period, relative to the intact control, while non-canopy ET increased. We attributed the elevated non-canopy ET in the girdled site each year to winter increases in sublimation, and summer increases in both soil evaporation and below-canopy transpiration. Although we expected that mortality of a canopy dominant would increase the availability of water and other resources to surviving vegetation, we observed a decrease in both soil volumetric water content and sap flow rates in the remaining trees at the girdled site, relative to the control. This post-girdling decrease in the performance of the remaining trees occurred during the severe 2011-2012 drought, suggesting that piñon mortality may trigger feedback mechanisms that leave PJ woodlands drier relative to undisturbed sites, and potentially more vulnerable to drought.

Morillas, L.↗

Study of the Osteoindictive Properties of Protein-Modified Polylactide Scaffolds

Bone marrow mesenchymal stromal cells are multipotent and can differentiate into cells of various tissues, which determines their high importance for clinical application. We performed an in vitro study of the osteogenic potential of mesenchymal stromal cells cultured on intact polylactide scaffolds or scaffolds modified with collagen I or fibrin. Scanning electron microscopy showed that the cells formed osteogenic nodules or osteogenic nodules on both intact and fibrin-modified polylactide scaffolds. Spectrophotometric detection of alkaline phosphatase activity on days 7 and 11 showed that mesenchymal stromal cell grown on intact polylactide scaffolds and on scaffolds modified with collagen type I or fibrin more intensively synthesized alkaline phosphatase than in the control (culture plastic). This dependence increases in the presence of osteogenic differentiation factors in the medium. After long-term culturing (4 weeks), the presence of calcium deposits detected by alizarin red staining confirmed the osteoinductive properties of intact and protein-modified polylactide scaffolds. These findings suggest that polylactide scaffolds and collagen I increase the osteogenic differentiation potential of mesenchymal stromal cells.

60 APPLIED LIFE SCIENCES↗

Calibration approach and range of observed sap flow influences transpiration estimates from thermal dissipation sensors

Calibrating thermal dissipation (TD) sap flow sensors has become increasingly important to accurately estimate whole-tree transpiration, but it is unclear how the calibration approach itself influences the resulting coefficients and estimates. Here, we compare the two most common calibration approaches, gravimetric and potometric, using TD sensors inserted into Eucalyptus benthamii tree stems. The gravimetric approach uses an excised stem segment devoid of branches and leaves and pushes water through the stem using gravity, a positive force. The potometric approach uses a severed stem containing an intact canopy placed upright in a reservoir where water is pulled through the stem via transpiration, a negative force. We hypothesized that the positive pressure associated with gravimetric calibration would overestimate conductive sapwood area relative to that estimated from potometric calibration and that coefficients from these different approaches would result in different estimates of transpiration when applied to intact trees. We also predicted that calibrations could improve transpiration estimates by targeting the range of observed sap flow rates (i.e., K values) in intact trees. Conductive sapwood area was higher under gravimetric calibrations and resulting estimates of transpiration were lower compared to potometric calibrations. Segmented calibration curves, which fit two separate curves for the relationship between sap flux density (Fd) and sap flux index (K) based on the range of sap flow rates observed in intact trees, increased transpiration estimates from both gravimetric and potometric coefficients and diminished the magnitude of difference in transpiration estimates between approaches. Researchers should be aware that calibration approach and range of observed sap flow profoundly influences transpiration estimates from TD sensors and this likely applies to calibrations of other heat-based sap flow sensors.

54 ENVIRONMENTAL SCIENCES↗

Aquaporin 0 plays a pivotal role in refractive index gradient development in mammalian eye lens to prevent spherical aberration

Highlights: • Intact AQP0 functions as fiber cell-to-fiber cell adhesion protein. • AQP0 facilitates reduction in extracellular space and lens water content. • AQP0 adhesion function aids in lens refractive index gradient (RING) formation. • AQP0 prevents lens spherical aberration by establishing RING. • AQP0 is critical for lens transparency and homeostasis. - Abstract: Aquaporin 0 (AQP0) is a transmembrane channel that constitutes ∼45% of the total membrane protein of the fiber cells in mammalian lens. It is critical for lens transparency and homeostasis as mutations and knockout cause autosomal dominant lens cataract. AQP0 functions as a water channel and as a cell-to-cell adhesion (CTCA) molecule in the lens. Our recent in vitro studies showed that the CTCA function of AQP0 could be crucial to establish lens refractive index gradient (RING). However, there is a lack of in vivo data to corroborate the role of AQP0 as a fiber CTCA molecule which is critical for creating lens RING. The present investigation is undertaken to gather in vivo evidence for the involvement of AQP0 in developing lens RING. Lenses of wild type (WT) mouse, AQP0 knockout (heterozygous, AQP0{sup +/−}) and AQP0 knockout lens transgenically expressing AQP1 (heterozygous AQP0{sup +/−}/AQP1{sup +/−}) mouse models were used for the study. Data on AQP0 protein profile of intact and N- and/or C-terminal cleaved AQP0 in the lens by MALDI-TOF mass spectrometry and SDS–PAGE revealed that outer cortex fiber cells have only intact AQP0 of ∼28 kDa, inner cortical and outer nuclear fiber cells have both intact and cleaved forms, and inner nuclear fiber cells have only cleaved forms (∼26–24 kDa). Knocking out of 50% of AQP0 protein caused light scattering, spherical aberration (SA) and cataract. Restoring the lost fiber cell membrane water permeability (P{sub f}) by transgene AQP1 did not reinstate complete lens transparency and the mouse lenses showed light scattering and SA. Transmission and scanning electron micrographs of lenses of both mouse models showed increased extracellular space between fiber cells. Water content determination study showed increase in water in the lenses of these mouse models. In summary, lens transparency, CTCA and compact packing of fiber cells were affected due to the loss of 50% AQP0 leading to larger extracellular space, more water content and SA, possibly due to alteration in RING. To our knowledge, this is the first report identifying the role of AQP0 in RING development to ward off lens SA during focusing.

60 APPLIED LIFE SCIENCES↗

Comparison of silver, cesium, and strontium release predictions using PARFUME with results from the AGR-1 irradiation experiment

Here, the PARFUME (PARticle FUel ModEl) code was used to predict the release of fission products silver, cesium, and strontium from tristructural isotropic coated fuel particles and compacts during the first irradiation experiment (AGR-1) of the Advanced Gas Reactor Fuel Development and Qualification program. The PARFUME model for the AGR-1 experiment used the fuel compact volume average temperature for each of the 620 days of irradiation to calculate the release of silver, cesium, and strontium from a representative particle for a select number of AGR-1 compacts. Post-irradiation examination measurements provided data on release of these fission products from fuel compacts and fuel particles, and retention of silver in the compacts outside of the silicon carbide (SiC) layer. PARFUME-predicted fractional release of silver, cesium, and strontium was determined and compared to the PIE measurements. For silver, comparisons show a trend of over-prediction at low burnup and under-prediction at high burnup. PARFUME has limitations in the modeling of the temporal and spatial distributions of the temperature and burnup across the compacts, which affects the accuracy of its predictions. Nevertheless, the comparisons on silver release lie in the same order of magnitude. Results show an overall over-prediction of the fractional release of cesium by PARFUME. For particles with failed SiC layers, the over-prediction is by a factor of up to 3, corresponding to a potential over-estimation of the diffusivity in uranium oxycarbide (UCO) by a factor of up to 250. For intact particles, whose release is much lower, the over-prediction is by a factor of up to 100, which could be attributed to an over-estimated diffusivity in SiC by about 40% on average. The release of strontium from intact particles is also over-predicted by PARFUME, which also points towards an over-estimated diffusivity of strontium in either SiC or UCO, or possibly both. The measured strontium fractional release from intact particles varied considerably from compact to compact, making it difficult to assess the effective over-estimation of the diffusivities. Moreover, the release of strontium from particles with failed SiC is difficult to observe experimentally due to the release from intact particles, preventing any conclusions to be made on the accuracy or validity of the PARFUME predictions and the modeled diffusivity of strontium in UCO.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

High tensile alloy of copper to mitigate current collector deformation in silicon electrodes for lithium-ion batteries

The volumetric changes of silicon electrodes, along with the strong adhesive properties of certain binders, can lead to plastic deformation of the current collector and create damage in the electrode coating. Here, in this study, we report a detailed study of silicon coatings on a high-tensile alloy (HTA) foil of copper with strength over twice that of conventional copper foils. The HTA current collectors with high mechanical strength can mitigate plastic deformation upon continuous cycling. At moderate areal capacities (2.5–3 mAh cm −2 ), conventional copper foils show significant wrinkling after only a few electrochemical cycles, whereas the HTA foils remain intact. We demonstrate viability of the HTA foils in large format xx6395 pouch cells, in which the HTA current collectors remain intact even at an areal capacity of 4.5 mAh cm −2 ; in contrast, wrinkles form in conventional copper current collectors increasing the likelihood of lithium plating. Computational studies show that stresses generated during cycling of silicon electrodes are very high in the current collector and at the current collector-coating interface, explaining the wrinkling of conventional Cu foils. Our studies highlight importance of current collector to solve the electrochemical and chemo-mechanical performance challenges associated with high-loading silicon electrodes.

Chemo-mechanical degradation↗

Irradiation performance of AGR-1 high temperature reactor fuel

The AGR-1 experiment contained 72 low-enriched uranium oxide/uranium carbide TRISO coated particle fuel compacts in six capsules irradiated to burnups of 11.2 to 19.6% FIMA, with zero TRISO coating failures detected during the irradiation. The irradiation performance of the fuel including the extent of fission product release and the evolution of kernel and coating microstructures was evaluated based on detailed examination of the irradiation capsules, the fuel compacts, and individual particles. Fractional release of 110m Ag from the fuel compacts was often significant, with capsule-average values ranging from 0.01 to 0.38. Analysis of silver release from individual compacts indicated that it was primarily dependent on fuel temperature history. Europium and strontium were released in small amounts through intact coatings, but were found to be significantly retained in the outer pyrocarbon and compact matrix. The capsule-average fractional release from the compacts was 1 × 10 –4 to 5 × 10 –4 for 154 Eu and 8 × 10 –7 to 3 × 10 –5 for 90 Sr. The average 134 Cs fractional release from compacts was <3 × 10 –6 when all particles maintained intact SiC. An estimated four particles out of 2.98 × 10 5 in the experiment experienced partial cesium release due to SiC failure during the irradiation, driving 134 Cs fractional release in two capsules to approximately 10 –5 . Identification and characterization of these particles has provided unprecedented insight into the nature and causes of SiC coating failure in high-quality TRISO fuel. In general, changes in coating morphology were found to be dominated by the behavior of the buffer and inner pyrolytic carbon (IPyC), and infrequently observed SiC layer damage was usually related to cracks in the IPyC. Palladium attack of the SiC layer was relatively minor, except for the particles that released cesium during irradiation, where SiC corrosion was found adjacent to IPyC cracks. In conclusion, palladium, silver, and uranium were found in the SiC layer of irradiated particles, and characterization of these elements within the SiC microstructure is the subject of ongoing focused study.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

Performance of AGR-1 high-temperature reactor fuel during post-irradiation heating tests

The fission product retention of irradiated low-enriched uranium oxide/uranium carbide tri-structural isotropic (TRISO) fuel compacts from the Advanced Gas-Cooled Reactor 1 (AGR-1) experiment has been evaluated at temperatures of 1600–1800 °C during post-irradiation safety tests. Fourteen compacts (a total of ~58,000 particles) with a burnup ranging from 13.4% to 19.1% fissions per initial metal atom (FIMA) have been tested using dedicated furnace systems at Idaho National Laboratory and Oak Ridge National Laboratory. The release of fission products 110mAg, 134Cs, 137Cs, 154Eu, 155Eu, 90Sr, and 85Kr was monitored while heating the fuel specimens in flowing helium. The behavior of silver, europium, and strontium appears to be dominated by inventory that was originally released through intact SiC coating layers during irradiation, but was retained in the compact at the end of irradiation and subsequently released during the safety tests. However, at a test temperature of 1800 °C, the data suggest that release of these elements through intact coatings may become significant after ~100 h. Cesium was very well retained by intact SiC layers, with a fractional release <5 × 10–6 after 300 h at 1600 °C or 100 h at 1800 °C. However, it was rapidly released from individual particles if the SiC layer failed, and therefore the overall cesium release fraction was dominated by the SiC defect and failure fractions in the fuel compacts. No complete TRISO coating layer failures were observed after 300 h at 1600 or 1700 °C, and 85 Kr release was very low during the tests (particles with failed SiC, but intact outer pyrocarbon, retained most of their krypton). Krypton release from TRISO failures was only observed after ~210 h at 1800 °C in one compact. As a result, post-safety-test examination of fuel compacts and particles has focused on identifying specific particles from each compact with notable fission product release and detailed analysis of the coating layers to understand particle behavior.

21 SPECIFIC NUCLEAR REACTORS AND ASSOCIATED PLANTS↗

Initial results from safety testing of US AGR-2 irradiation test fuel

Two cylindrical compacts containing tristructural isotropic (TRISO)-coated particles with kernels that contained a mixture of uranium carbide and uranium oxide (UCO) and two compacts with UO 2 -kernel TRISO particles have undergone 1600°C safety testing. These compacts were irradiated in the US Advanced Gas Reactor Fuel Development and Qualification Program's second irradiation test (AGR-2). The time-dependent releases of several radioisotopes ( 110m Ag, 134 Cs, 137 Cs, 154 Eu, 155 Eu, 90 Sr, and 85 Kr) were monitored while heating the fuel specimens to 1600°C in flowing helium for 300 h. The UCO compacts behaved similarly to previously reported 1600°C-safety-tested UCO compacts from the AGR-1 irradiation. No failed TRISO or failed SiC were detected (based on krypton and cesium release), and cesium release through intact SiC was very low. Release behavior of silver, europium, and strontium appeared to be dominated by inventory originally released through intact coating layers during irradiation but retained in the compact matrix until it was released during safety testing. Both UO 2 compacts exhibited cesium release from multiple particles whose SiC failed during the safety test. Europium and strontium release from these two UO 2 compacts appeared to be dominated by release from the particles with failed SiC. Silver release was characteristically like the release from the UCO compacts in that an initial release of the majority of silver trapped in the matrix occurred during ramping to 1600°C. However, additional silver release was observed later in the safety testing due to the UO 2 TRISO with failed SiC. Failure of the SiC layer in the UO 2 fuel appears to have been dominated by CO corrosion, as opposed to the palladium degradation observed in AGR-1 UCO fuel.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

Selenium Preferentially Accumulates in the Eye Lens Following Embryonic Exposure: A Confocal X-ray Fluorescence Imaging Study

Maternal transfer of elevated selenium (Se) to offspring is an important route of Se exposure for fish in the natural environment. However, there is a lack of information on the tissue specific spatial distribution and speciation of Se in the early developmental stages of fish, which provide important information about Se toxicokinetics. The effect of maternal transfer of Se was studied by feeding adult zebrafish a Se-elevated or a control diet followed by collection of larvae from both groups. Novel confocal synchrotron-based techniques were used to investigate Se within intact preserved larvae. Confocal X-ray fluorescence imaging was used to compare Se distributions within specific planes of an intact larva from each of the two groups. The elevated Se treatment showed substantially higher Se levels than the control; Se preferentially accumulated to highest levels in the eye lens, with lower levels in the retina, yolk and other tissues. Confocal X-ray absorption spectroscopy was used to determine that the speciation of Se within the eye lens of the intact larva was a selenomethionine-like species. Preferential accumulation of Se in the eye lens may suggest a direct cause-and-effect relationship between exposure to elevated Se and Se-induced ocular impairments reported previously. This study illustrates the effectiveness of confocal X-ray fluorescence methods for investigating trace element distribution and speciation in intact biological specimens

Choudhury, Sanjukta↗

A Targeted Mass Spectrometric Assay for Reliable Sensitive Hepcidin Quantification

Hepcidin, a cysteine-rich peptide hormone, secreted mainly by the liver, plays a central role in iron metabolism regulation. Emerging evidence suggests that disordered iron metabolism is a risk factor for various types of diseases including cancers. However, it remains challenging to apply current mass spectrometry (MS)-based hepcidin assays for precise quantification due to the low fragmentation efficiency of intact hepcidin as well as synthesis difficulties for the intact hepcidin standard. To address these issues we recently developed a reliable sensitive targeted MS assay for hepcidin quantification that uses fully alkylated rather than intact hepcidin as the internal standard. Limits of detection and quantification were determined to be <0.5 ng/mL and 1 ng/mL, respectively, which are better than those from currently available MS-based hepcidin assays. Application of the alkylated hepcidin assay to 70 clinical plasma samples (42 normal and 28 ovarian cancer patient samples) enabled reliable detection of endogenous hepcidin from the plasma samples, as well as conditioned culture media. The hepcidin concentrations ranged from 0.0 to 95.6 ng/mL across normal and cancer plasma specimens. Interestingly, cancer patients were found to have significantly higher hepcidin concentrations compared to normal patients (mean: 20.6 ng/ml for cancer; 5.94 ng/ml for normal) with the p value of <0.001. Our results demonstrate that the newly developed hepcidin assay has better sensitivity and quantification accuracy than current MS-based hepcidin assays without the challenges in synthesis of intact hepcidin standard and accurately determining its absolute amount.

59 BASIC BIOLOGICAL SCIENCES↗

Top-down mass spectrometry and assigning internal fragments for determining disulfide bond positions in proteins

Disulfide bonds in proteins have a substantial impact on protein structure, stability, and biological activity. Localizing disulfide bonds is critical for understanding protein folding and higher-order structure. Conventional top-down mass spectrometry (TD-MS), where only terminal fragments are assigned for disulfide-intact proteins, can access disulfide information, but suffers from low fragmentation efficiency, thereby limiting sequence coverage. Here, we show that assigning internal fragments generated from TD-MS enhances the sequence coverage of disulfide-intact proteins by 20–60% by returning information from the interior of the protein sequence, which cannot be obtained by terminal fragments alone. Further, the inclusion of internal fragments can extend the sequence information of disulfide-intact proteins to near complete sequence coverage. Importantly, the enhanced sequence information that arise from the assignment of internal fragments can be used to determine the relative position of disulfide bonds and the exact disulfide connectivity between cysteines. The data presented here demonstrates the benefits of incorporating internal fragment analysis into the TD-MS workflow for analyzing disulfide-intact proteins, which would be valuable for characterizing biotherapeutic proteins such as monoclonal antibodies and antibody–drug conjugates.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Isotopic insights into methane production, oxidation, and emissions in Arctic polygon tundra

Arctic wetlands are currently net sources of atmospheric CH 4 . Due to their complex biogeochemical controls and high spatial and temporal variability, current net CH 4 emissions and gross CH 4 processes have been difficult to quantify, and their predicted responses to climate change remain uncertain. Here, we investigated CH 4 production, oxidation, and surface emissions in Arctic polygon tundra, across a wet-to-dry permafrost degradation gradient from low-centered (intact) to flat- and high-centered (degraded) polygons. From 3 microtopographic positions (polygon centers, rims, and troughs) along the permafrost degradation gradient, we measured surface CH 4 and CO 2 fluxes, concentrations and stable isotope compositions of CH 4 and DIC at three depths in the soil, and soil moisture and temperature. More degraded sites had lower CH 4 emissions, a different primary methanogenic pathway, and greater CH 4 oxidation than did intact permafrost sites, to a greater degree than soil moisture or temperature could explain. Surface CH 4 flux decreased from 64 nmol m -2 s -1 in intact polygons to 7 nmol m -2 s -1 in degraded polygons, and stable isotope signatures of CH 4 and DIC showed that acetate cleavage dominated CH 4 production in low-centered polygons, while CO 2 reduction was the primary pathway in degraded polygons. We see evidence that differences in water flow and vegetation between intact and degraded polygons contributed to these observations. In contrast to many previous studies, these findings document a mechanism whereby permafrost degradation can lead to local decreases in tundra CH 4 emissions.

54 ENVIRONMENTAL SCIENCES↗

Soil disturbance and invasion magnify CO 2 effects on grassland productivity, reducing diversity

Climate change, disturbance, and plant invasion threaten grassland ecosystems, but their combined and interactive effects are poorly understood. Here, in this study, we examine how the combination of disturbance and plant invasion influences the sensitivity of mixed-grass prairie to elevated carbon dioxide (eCO 2 ) and warming. We established subplots of intact prairie and disturbed/invaded prairie within a free-air CO 2 enrichment (to 600 ppmv) by infrared warming (+1.5°C day, 3°C night) experiment and followed plant and soil responses for 5 years. Elevated CO 2 initially led to moderate increases in biomass and plant diversity in both intact and disturbed/invaded prairie, but these effects shifted due to strong eCO 2 responses of the invasive forb Centaurea diffusa. In the final 3 years, biomass responses to eCO 2 in disturbed/invaded prairie were 10 times as large as those in intact prairie (+186% vs. +18%), resulting in reduced rather than increased plant diversity (-17% vs. +10%). At the same time, warming interacted with disturbance/invasion and year, reducing the rate of topsoil carbon recovery following disturbance. The strength of these interactions demonstrates the need to incorporate disturbance into predictions of climate change effects. In contrast to expectations from studies in intact ecosystems, eCO 2 may threaten plant diversity in ecosystems subject to soil disturbance and invasion.

54 ENVIRONMENTAL SCIENCES↗

Laser‐ablation electrospray ionization mass spectrometry with ion mobility separation reveals metabolites in the symbiotic interactions of soybean roots and rhizobia

Summary Technologies enabling in situ metabolic profiling of living plant systems are invaluable for understanding physiological processes and could be used for rapid phenotypic screening (e.g., to produce plants with superior biological nitrogen‐fixing ability). The symbiotic interaction between legumes and nitrogen‐fixing soil bacteria results in a specialized plant organ (i.e., root nodule) where the exchange of nutrients between host and endosymbiont occurs. Laser‐ablation electrospray ionization mass spectrometry ( LAESI ‐ MS ) is a method that can be performed under ambient conditions requiring minimal sample preparation. Here, we employed LAESI ‐ MS to explore the well characterized symbiosis between soybean ( Glycine max L. Merr.) and its compatible symbiont, Bradyrhizobium japonicum . The utilization of ion mobility separation ( IMS ) improved the molecular coverage, selectivity, and identification of the detected biomolecules. Specifically, incorporation of IMS resulted in an increase of 153 differentially abundant spectral features in the nodule samples. The data presented demonstrate the advantages of using LAESI – IMS – MS for the rapid analysis of intact root nodules, uninfected root segments, and free‐living rhizobia. Untargeted pathway analysis revealed several metabolic processes within the nodule (e.g., zeatin, riboflavin, and purine synthesis). Compounds specific to the uninfected root and bacteria were also detected. Lastly, we performed depth profiling of intact nodules to reveal the location of metabolites to the cortex and inside the infected region, and lateral profiling of sectioned nodules confirmed these molecular distributions. Our results established the feasibility of LAESI – IMS – MS for the analysis and spatial mapping of plant tissues, with its specific demonstration to improve our understanding of the soybean‐rhizobial symbiosis.

Stopka, Sylwia A.↗